Efficient DNA transfection mediated by the C-terminal domain of human immunodeficiency virus type 1 viral protein R

被引:56
作者
Kichler, A
Pages, JC
Leborgne, C
Druillennec, S
Lenoir, C
Coulaud, D
Delain, E
Le Cam, E
Roques, BP
Danos, O
机构
[1] Genethon III, CNRS URA 1923, F-91002 Evry, France
[2] CNRS UMR 8600, INSERM U266, Dept Pharmacochim Mol & Struct, Paris, France
[3] Inst Gustave Roussy, CNRS, UMR 8532, Lab Microscopie Mol & Cellulaire, Villejuif, France
关键词
D O I
10.1128/JVI.74.12.5424-5431.2000
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Viral protein R (Vpr) of human immunodeficiency virus type I is produced late in the virus life cycle and is assembled into the virion through binding to the Gag protein. It is known to play a significant role early in the viral life cycle by facilitating the nuclear import of the preintegration complex in nondividing cells. Vpr is also able to interact with nucleic acids, and we show here that it induces condensation of plasmid DNA. We have explored the possibility of using these properties in DNA transfection experiments. We report that the C-terminal half of the protein (Vpr(52-96)) mediates DNA transfection in a variety of human and nonhuman cell lines,with efficiencies comparable to those of the best-known transfection agents. Compared with polylysine, a standard polycationic transfection reagent, Vpr(52-96) was 10- to 1,000-fold more active, Vpr(52-96)-DNA complexes were able to reach the cell nucleus through a pH-independent mechanism. These observations possibly identify an alternate pathway for DNA transfection.
引用
收藏
页码:5424 / 5431
页数:8
相关论文
共 56 条
[1]   The human immunodeficiency virus type 1 Vpr transactivator: Cooperation with promoter-bound activator domains and binding to TFIIB [J].
Agostini, I ;
Navarro, JM ;
Rey, F ;
Bouhamdan, M ;
Spire, B ;
Vigne, R ;
Sire, J .
JOURNAL OF MOLECULAR BIOLOGY, 1996, 261 (05) :599-606
[2]   A VERSATILE VECTOR FOR GENE AND OLIGONUCLEOTIDE TRANSFER INTO CELLS IN CULTURE AND IN-VIVO - POLYETHYLENIMINE [J].
BOUSSIF, O ;
LEZOUALCH, F ;
ZANTA, MA ;
MERGNY, MD ;
SCHERMAN, D ;
DEMENEIX, B ;
BEHR, JP .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (16) :7297-7301
[3]  
Boussif O, 1996, GENE THER, V3, P1074
[4]   BAFILOMYCINS - A CLASS OF INHIBITORS OF MEMBRANE ATPASES FROM MICROORGANISMS, ANIMAL-CELLS, AND PLANT-CELLS [J].
BOWMAN, EJ ;
SIEBERS, A ;
ALTENDORF, K .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (21) :7972-7976
[5]  
Bukrinsky M, 1999, REV MED VIROL, V9, P39, DOI 10.1002/(SICI)1099-1654(199901/03)9:1<39::AID-RMV235>3.0.CO
[6]  
2-3
[7]   HIGH-EFFICIENCY TRANSFORMATION BY DIRECT MICRO-INJECTION OF DNA INTO CULTURED MAMMALIAN-CELLS [J].
CAPECCHI, MR .
CELL, 1980, 22 (02) :479-488
[8]   ENHANCED RESISTANCE TO NUCLEASE DEGRADATION OF NUCLEIC-ACIDS COMPLEXED TO ASIALOGLYCOPROTEIN-POLYLYSINE CARRIERS [J].
CHIOU, HC ;
TANGCO, MV ;
LEVINE, SM ;
ROBERTSON, D ;
KORMIS, K ;
WU, CH ;
WU, GY .
NUCLEIC ACIDS RESEARCH, 1994, 22 (24) :5439-5446
[9]  
COHEN EA, 1990, J ACQ IMMUN DEF SYND, V3, P11
[10]   VPR IS REQUIRED FOR EFFICIENT REPLICATION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 IN MONONUCLEAR PHAGOCYTES [J].
CONNOR, RI ;
CHEN, BK ;
CHOE, S ;
LANDAU, NR .
VIROLOGY, 1995, 206 (02) :935-944